| Skeletal muscle genes are important potentially functional candidate genes for livestock production and meat quality. Myosin regulatory light chain (MLC) regulates myofilament activation via phosphorylation by Ca2+Dependent myosin light chain kinase, myosin light chain, phosphorylatable, fast skeletal muscle(MYLPF) gene as one of myosin light chain gene has important effects on livestock meat quality. Human and animal fat metabolism is regulated by a varies of factors and the most important factor is the genetic factor. Phosphotyrosine interaction domain1(PID1) is a new gene discovered in recent years, which has a direct role in the cell growth and lipid process as a signaling molecule, over-expresses in the fat cells directly leaded to produce obesity phenotype.The cDNA of the MYLPF and PID1from the longissimus dorsi and abdominal muscle of Tianfu goat was cloned and sequenced. The results showed that MYLPF full-length coding sequence consists of513bp and encodes170amino acids, it was then deposited into the GenBank database (GenBank accession No. JN107562). Six phosphorylation sites were predicted in the translated PID1protein. Two EF-hand superfamily domain of MYLPF gene conserved between Tianfu goat and other animals. The deduced amino acid sequence of MYLPF shared significant identity with the MYLPF from other mammals. A phylogenetic tree analysis revealed that the Tianfu goat MYLPF protein has a close genetic relationship and evolutional distance with MYLPF in other mammals. The results showed that the full sequence of the Tianfu goat PID1cDNA was896bp long and contained a654bp long coding region that encoded a217amino acid sequence, it was then deposited into the GenBank database (GenBank accession No. JN257257). Fifteen phosphorylation sites were predicted in the translated PID1protein. The protein had a phosphotyrosine-binding domain between Arg53and Ile199. A phylogenic tree based on the PID1proteins from other species revealed that the Tianfu goat protein was closely related to cattle PID1.Analysis by RT-PCR showed that the MYLPF mRNA was detected in heart, liver, spleen, lungs, kidney, gastrocnemius, abdominal muscle and longissimus dorsi. In particular, high expression levels of MYLPF mRNA were detected in the longissimus dorsi. gastrocnemius and abdominal muscle, and low level of expressions were observed in liver, spleen, lungs and kidney. In addition, the temporal expression analysis further showed MYLPF expression decreased gradually with age in the skeletal muscle. Fluorescence quantitative PCR analyses revealed that PID1was expressed in the8tissues of Tianfu goat. In particular, high expression levels of PID1were detected in liver and abdominal muscle, and low expression levels were seen in lungs. Furthermore, the PID1mRNA expression levels in the longissimus dorsi muscles increased gradually with the age of the Tianfu goat.In this study, we used the Western blotting method to detect MYLPF and PID1protein expression levels in eight tissues from3and12-month-old goat. The MYLPF protein in four of the tissues in which MYLPF was shown to be expressed; the four exceptions were liver, spleen, lungs and kidney. The PID1protein in six of the tissues in which PID1was shown to be expressed; the two exceptions were liver and spleen. But in heart and kidney that were equally abundant but had significantly bigger molecular weights than PID1.The IMF content increased gradually with age of from3to12months the Tianfu goat in the longissimus dorsi muscles, and reached a peak in12months. Gene expression level and IMF content correlation analysis showed that:The expression of MYLPF and the IMF content showed negative correlation, and the correlation coefficient was-0.925; The expression of PID1gene and the IMF content showed a highly significant positive correlation, and the correlation coefficient was0.943;... |
